Methods for simultaneous and quantitative isolation of mitochondrial DNA, nuclear DNA and RNA from mammalian cells

Biotechniques. 2020 Dec;69(6):436-442. doi: 10.2144/btn-2020-0114. Epub 2020 Oct 26.

Abstract

The aim of this study was to assess two protocols for their capacities to simultaneously isolate RNA, mtDNA and ncDNA from mammalian cells. We compared the Invitrogen TRIzol-based method and Qiagen DNeasy columns, using the HepG2 cell line and human primary glioblastoma stem cells. Both methods allowed the isolation of all three types of nucleic acids and provided similar yields in mtDNA. However, the yield in ncDNA was more than tenfold higher on columns, as observed for both cell types. Conversely, the TRIzol method proved more reproducible and was the method of choice for isolating RNA from glioblastoma cells, as demonstrated for the housekeeping genes RPLP0 and RPS9.

Keywords: RNA; mammalian cells; mitochondria; mitochondrial DNA (mtDNA); nuclear DNA (ncDNA).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biochemistry / methods*
  • Cell Nucleus / metabolism*
  • DNA, Mitochondrial / isolation & purification*
  • Glioblastoma / metabolism
  • Glioblastoma / pathology
  • Hep G2 Cells
  • Humans
  • Mammals / metabolism*
  • Neoplastic Stem Cells / metabolism
  • RNA / isolation & purification*
  • RNA, Messenger / isolation & purification
  • Reagent Kits, Diagnostic

Substances

  • DNA, Mitochondrial
  • RNA, Messenger
  • Reagent Kits, Diagnostic
  • RNA